Collaborative Research: Maximizing Therapeutic DNA Process Productivity

DNA疫苗作为一种增强免疫力的方式正变得越来越重要。归根结底，DNA疫苗应该比传统疫苗更安全、更有效，然而，目前我们生产必要DNA的能力存在限制。为了满足对DNA疫苗和其他DNA产品的需求，提高生产过程的生产率将是有益的。在降低成本的同时，更快的处理也将使DNA疫苗生产商能够更快地对病原体突变和疾病爆发做出反应。拟议的工作解决了这个问题。其主要思想是，当宿主和质粒DNA同时进行工程时，DNA产量显著增加，而宿主在简单而廉价的培养基上生长迅速。研究人员将结合代谢工程、蛋白质组学、核磁共振、生物处理和数学建模来实现他们的目标。这项工作将把代谢工程实践从目前关注的小分子和蛋白质扩展到DNA产品，这对推进代谢工程领域具有重要意义。从更根本的角度来看，目前还不太清楚的质粒DNA合成与宿主细胞代谢的整合将会得到改善。联合突变在控制质粒复制中的作用也还没有得到很好的研究。这项工作将揭示这些效应的协同程度，这将是结构分子生物学家感兴趣的。氧化还原辅因子平衡的影响也将被阐明，这是目前正在进行的代谢工程努力的兴趣所在。成功的结果将加快治疗性DNA产品的生产，并促进临床试验。这一合作项目将为涉及代谢工程、计算、核磁共振、蛋白质组学和DNA产品生物加工的研究生和本科生(包括少数族裔学生)提供广泛的教育和培训。

0967874DomachDNA vaccines are becoming increasingly important as a way to confer immunity. Ultimately, DNA vaccines should be safer and more effective than traditional vaccines, however, currently, there are limitations in our ability to produce the necessary DNA. To meet the demand for DNA vaccines and other DNA products, it would be beneficial to increase the productivity of production processes. While reducing cost, faster processing would also enable DNA vaccine producers to more quickly respond to pathogen mutations and disease outbreaks. The proposed work addresses this problem. The main idea is when host and plasmid DNA are concurrently engineered, DNA production increases dramatically, while the host grows quickly on simple and inexpensive medium. The investigators will use a combination of metabolic engineering, proteomics, NMR, bioprocessing, and mathematical modeling to achieve their goals. The work will extend metabolic engineering practice from the current foci of small molecules and proteins to DNA products, which is important in advancing the field of metabolic engineering. From a more fundamental standpoint, the integration of plasmid DNA synthesis with host cell metabolism, which is not now well understood, will be improved. The effect of combining mutations in the control of plasmid replication is also not yet well explored. The work will reveal the extent to which the effects are synergistic, which would be of interest to structural molecular biologists. The impact of redox cofactor balancing will also be elucidated, which is of current interest to a number of ongoing metabolic engineering efforts. A successful outcome will accelerate the production of therapeutic DNA products and facilitate clinical trials. This collaborative project would provide broad education and training for the graduate and undergraduate students (including minority students) involved in metabolic engineering, computation, NMR, proteomics, and the bioprocessing of DNA products.